1. Field of the Invention
The present invention relates to a method and an apparatus for analyzing organic macromolecular components in a solution based on a flow analysis method, and relates to applications thereof. In particular, the present invention relates to an analytical method and an apparatus suitable for quantitatively analyzing very small amounts of glues or gelatins contained in metallic electrolytic solutions such as copper electrolytic solutions and plating solutions.
2. Discussion of the Background
Additives are added to electrolytic solutions and plating solutions for various purposes, such as improving brightness and smoothness of electrodeposited metal surfaces, hardening plated layers, etc. Generally, glue additives are used in electrolytic smelting, and gelatins are used in plating. Herein, for quality control, it is very important to keep concentrations thereof within specified ranges. For example, although glues are used for improving the smoothness of electrodeposited surfaces, when concentrations thereof are too high polarization is increased to such a great degree that there are problems with deposition of impurities such as bismuth contained in the electrolytic solutions, etc.
Therefore, a method for analyzing proteins, such as glues or gelatins contained in solutions, with a high degree of precision has been required. Most of the conventional methods for analyzing proteins are performed under conditions being from weakly acidic to weakly basic, and there are few analytical methods which can be applied under strongly acidic conditions of a pH 1 or less, as in electrolytic solutions, plating solutions, etc. Hitherto, proteins such as glues or gelatins contained in electrolytic solutions, plating solutions, etc., have been measured by a potentiometric titration, the Kjeldahl method, etc., although specific apparatuses and complicated operations have been required in those cases. In the Kjeldahl method, glues are measured after the nitrogen in the glues is converted into ammonia, although in many cases nitrogen compounds other than proteins are contained in the electrolytic solutions so as to make precise measurements difficult.
In addition, as a quantitative method for analyzing gelatins and glues in a strongly acidic solution, a method in which gelatins and glues are collected on membrane filters, are bonded with a specific reagent (Amide Black 10B coloring matter), and after excess coloring, matters are washed out, the coloring matters are eluted so as to determine gelatins, etc., (Japanese Unexamined Patent Application Publication No. 2-69660); and a method in which after gelatins, etc., are collected on a filter, the resulting filter is dried so as to determine gelatins, etc., based on reflectance (Japanese Unexamined Patent Application Publication No. 6-337247), etc., are reported. Regarding these methods, there are, however, problems in that the amount of collected glues is greatly affected by pore diameters of the filter so as to increase measurement errors. A filtering apparatus for collecting with a filter and complicated operations therefor are required, and the analytical cost is increased due to the disposable filters.
The present invention has solved the aforementioned problems in the conventional analytical methods. Accordingly, the present invention provides a flow analysis method and an apparatus by which organic macromolecular components such as glues and gelatins in solutions can be promptly and precisely quantified even in strongly acidic solutions.
The present invention resides in the completion of a measurement system with a high degree of reliability and with handling ease, in which the operations of separating an organic macromolecular component in a solution by adsorption on a resin, leading this to a gel permeation chromatograph, and analysis are performed based on the flow analysis.
That is, the present invention relates to the following flow analysis methods for separating and analyzing an organic macromolecular component in a sample.
According to a first aspect of the present invention, a method for analyzing an organic macromolecular component based on a flow analysis method with a measurement system including a sample introduction section, a preparation section, and a measuring section, includes the steps of supplying a sample together with a carrier solution into the measurement system through the introduction section, leading the sample to the preparation section and separating the organic macromolecular component in the sample, and leading the separated organic macromolecular component to the measuring section for analysis. Herein, the step of separating the organic macromolecular component includes the steps of adsorbing the organic macromolecular component in the sample on a resin and introducing an eluant solution into the preparation section and eluting the organic macromolecular component adsorbed on the resin.
Preferably, the method of analysis may further include the steps of supplying the sample into the measurement system through the introduction section while a pipeline for feeding the sample running from the introduction section to the preparation section and a pipeline for discharging the solution running from the preparation section to the outside of the measurement system are connected to the preparation section; and leading the sample to the preparation section, where the organic macromolecular component in the sample is adsorbed on the resin in the preparation section, and leading the discharged solution from the preparation section to the outside of the measurement system. Switchable pipelines are connected to the preparation section so that a pipeline for supplying the eluant solution and a pipeline connected to the measuring section are connected to the preparation section, introducing the eluant solution into the preparation section so as to elute the organic macromolecular component adsorbed on the resin, and leading the resulting solution containing the organic macromolecular component to the measuring section for analysis.
Preferably, the method of analysis may further include the steps in which the eluant solution is used after a buffer solution to prevent the organic macromolecular components from coagulating is added thereto, or the buffer solution is added to the solution containing the organic macromolecular component between the preparation section and the measuring section.
Preferably, the method of analysis may further include the step in which a gel permeation chromatography is used as an analyzing device so as to fractionate and analyze the organic macromolecular component separated from the sample.
Preferably, the method of analysis may further include the step in which an amount of glues or gelatins is separated from the sample, or amounts of decomposition products thereof are measured.
Preferably, the method of analysis may further include the step in which a gel permeation chromatography is used as an analyzing device in the measuring section so as to fractionate glues or gelatins separated from the sample and to measure the amount thereof or the amounts of decomposition products thereof.
Preferably, the method of analysis may further include the step in which an acid-proof and hydrophobic adsorption resin is used as a resin for adsorbing the organic macromolecular component in the sample.
Preferably, the method of analysis may further include the step in which an electrolytic solution taken from a step of metallic electrolysis or a plating solution taken from a step of plating is used as a sample solution.
Preferably, the method for controlling a metallic electrolysis process with the method of analysis may further include the steps of taking a sample solution from an electrolytic solution in the step of metallic electrolysis, measuring the amount of glues or gelatins separated from the sample of the electrolytic solution or amounts of decomposition products thereof, and feeding the results of the measurement back to the step of metallic electrolysis.
Preferably, the method for controlling a plating process with the method of analysis may further include the steps of taking a sample solution from a plating solution in the step of plating, measuring the amount of glues or gelatins separated from the sample of the plating solution or amounts of decomposition products thereof, and feeding the results of the measurement back to the step of plating.
In the method of analysis according to the present invention, as described above, a series of the operations of separating proteins and organic macromolecular components such as glues and gelatins contained in the sample by adsorption on the resin from the sample, are lead to a gel permeation chromatograph, where analysis is performed based on the flow analysis method. Since the operations from the introduction of the sample to the analysis of the organic macromolecular component can be continuously performed in a short time, the analytical results can be promptly obtained. Therefore, regarding the organic macromolecular component, behaviors during decomposition and intermediate products can be grasped. Since the separation is performed using the hydrophobic adsorption resin and the analysis is performed using the gel permeation chromatography, the organic macromolecular component can be analyzed independent of the molecular weight. Furthermore, when the organic macromolecular components are fed to the measuring section, a buffer solution for preventing the coagulation thereof may be added so as not to cause a blockage of the pipeline, etc. Therefore, the analysis can be performed with a high degree of reliability. This buffer solution may be blended beforehand with the eluant solution or may be added between the preparation section and the measuring section. When the buffer solution is added after the organic macromolecular components adsorbed on the resin are eluted, the effect of eluting is improved and the effect of preventing the organic macromolecular components from coagulating is also improved.
The flow analysis method according to the present invention can be applied to strongly acidic solutions such as metallic electrolytic solutions and plating solutions by using an acid-proof and hydrophobic adsorption resin as the resin for adsorbing the organic macromolecular components. Therefore, the quantitative analysis of glues contained in the electrolytic solutions of copper electrolytic smelting, etc., can be easily performed so that it can be used as a method for controlling electrolytic operations according to the method of analysis of the present invention. Since a continuous automatic analysis is possible instead of a conventional batch method for controlling electrolysis by manual work, long term and accurate operation control of the electrolytic smelting is possible.
The present invention further relates to the following apparatuses for flow analysis.
According to a second aspect of the present invention, an apparatus for flow analysis of an organic macromolecular component includes a sample introduction section, a preparation section, and a measuring section integrally connected by pipelines, in which the preparation section is provided with an adsorbing device for adsorbing the organic macromolecular component and an eluting device for separating the organic macromolecular component in the sample, and the measuring section is provided with a fractionating and analyzing device for fractionating and analyzing the separated organic macromolecular component.
Preferably, the apparatus for flow analysis may further include a column filled with a resin for adsorbing the organic macromolecular component in the sample, and a pipeline for introduction, running from the introduction section to the outside of the measurement system, and a pipeline for elution, running from a supply source of the elution solution to the measuring section, where each is connected to the column so as to be freely switched from each other, the adsorbing device and the eluting device for the organic macromolecular component being composed thereof.
Preferably, the apparatus for flow analysis may further include an acid-proof and hydrophobic adsorption resin which is used as a resin for adsorbing the organic macromolecular component in the sample, and the protein is separated using a column filled with the aforementioned resin.
Preferably, the apparatus for flow analysis may further include a buffering section which is between the preparation section and the measuring section so as to add a buffer solution for preventing the organic macromolecular components from coagulating.
Preferably, the apparatus for flow analysis may further include a gel permeation chromatograph in the measuring section as an analytical device for measuring the organic macromolecular component.
Preferably, the apparatus for flow analysis may further include a column filled with a hydrophilic polymer gel having an exclusion molecular weight limit of 5xc3x97103 or more in the gel permeation chromatograph so that the molecular weight fractionation of the proteins is performed using the hydrophilic polymer gel.
Preferably, the apparatus for flow analysis may further include columns in a plurality of stages, each column being filled with a hydrophilic polymer gel having a different exclusion molecular weight limit.
Preferably, the apparatus for flow analysis may further include a thermostatic chamber in the measuring section, and may be further provided with a gel permeation chromatograph in the thermostatic chamber.
Preferably, the apparatus for flow analysis may further include sulfuric acid, hydrochloric acid, or nitric acid each having a concentration of 0.1 M or less, or a mixed acid solution thereof which is used as a carrier solution.
Preferably, the apparatus for flow analysis may further include 20% to 40% of methanol, ethanol, other lower alcohols, or 20% to 40% of acetonitrile which is used as the eluant solution.
Preferably, the apparatus for flow analysis may further include a mixture solution of the eluant solution and a phosphoric acid buffer solution for preventing organic macromolecular components from coagulating, which is introduced into a detecting device in the measuring section.
Preferably, the apparatus for flow analysis may further include a flow rate of the phosphoric acid solution which is introduced into the detecting device in the measuring section as 1 mL/min or less.
Preferably, the apparatus for flow analysis may further include pipelines which are made of tubes 1 mm or less in inner diameter, and made of stainless steel, TEFZEL (i.e., ethylene-tetrafluoroethylene), or PEEK.
Preferably, the apparatus for flow analysis may further include an automatic control device for controlling the feed of the solution and the discharge of the solution in the sample introduction section and in the preparation section, for temperature adjustment in the measuring section, and for the actions of a detecting section so as to continuously and automatically perform operations from sample introduction to fractionation, and analysis by way of separation of the organic macromolecular component.
According to the aforementioned apparatus for flow analysis, the organic macromolecular components contained in the sample can be separated with operational ease and can be analyzed. Furthermore, the column for adsorbing the organic macromolecular components, and the pipelines for sample introduction and for leading the separated organic macromolecular component to the measuring section, each connected to the resin column so as to be freely switched from each other, are provided. Therefore, feed of the sample to the resin column and discharge, adsorption of the organic macromolecular component by the resin and elution, and feed of the solution to the measuring section can be mechanically and smoothly performed in a short time. By the buffering section being provided between the preparation section and the measuring section, separated organic macromolecular components are prevented from coagulating so as not to cause a blockage of the pipeline and the analysis can be performed with a high degree of reliability.
Furthermore, since a measurement system is made so that the acid-proof and hydrophobic adsorption resin is used as the resin for adsorbing the organic macromolecular component in the sample, the gel permeation chromatography is used as the analytical device in the measuring section, the column filled with the hydrophilic polymer gel having the exclusion molecular weight limit of 5xc3x97103 or more is preferably used for the gel permeation chromatography, and if necessary, columns are connected in a plurality of stages, each column being filled with a hydrophilic polymer gel having a different exclusion molecular weight limit, precise analysis can be performed in response to the molecular weight of the organic macromolecular component. The aforementioned columns are preferably provided in a thermostatic chamber.
Since sulfuric acid, hydrochloric acid, or nitric acid each having a concentration of 0.1 M or less, or a mixed acid solution thereof is used as a carrier solution, 40% or less of methanol, ethanol, other lower alcohols, or 40% or less of acetonitrile is used as the eluant solution, and preferably, a mixture solution of the eluant solution and a phosphoric acid buffer solution for preventing organic macromolecular components from coagulating is used, the proteins can be precisely and smoothly analyzed.
In the apparatus for analysis according to the present invention, since the measurement system of the sample introduction section to the measuring section by way of the preparation section is integrally communicated by pipelines, and pipelines connected to the preparation section are made so as to be freely switched from each other, the operations of feeding the solution and discharging the solution in each section are able to be automatically controlled. Therefore, a series of operations from the introduction of the sample to the analysis can be automated with the aforementioned automatic control device being provided.